How proteomics shed light in understanding host-parasite interplay and clinical consequences during trypanosome infectious process

How
proteomics
shed
light
in
understanding
host‐parasite
interplay



and
clinical
consequences
during
trypanosome
infec8ous
process


P.
Holzmuller

_{,
P.
Grébaut}

_{,
N.
Parra‐Gimenez}

_{,
J.P.
Brizard}

_{,
E.
Deme;re}

_{,
M.
Seveno}

_{,
M.
Gonza?}

_{,
G.
Cuny}


 
Plate‐forme
DIGE,
Montpellier,
France
 
 
 
 Plate‐forme
de
protéomique
foncNonnelle,
Montpellier,
France


Secreted


protein
 Trypanosoma
species
 Pep8des/%cover/Mowse
score
 Propsearch
server
data
 Hypothe8cal
func8on
 Link
with
clinical
disorders



V1
 T.
congolense
 6
/
36
/
276
 Serine
protease
inhibior


2.4
precursor

 B‐SERPIN
B9

inhibitor/régulator
of
granzymes[3]


,
protecFon
of
trypanosomes
from
T
 cytotoxic
lymphocytes
acFvated
by
 TLTF[4]
 Immuno‐suppression,
 Trypanosome
development
 T8
 T.
congolense
 4
/
21
/
132
 EukaryoNc
translaNon


iniNaNon
factor
5

 eIF‐5
dicrease
protein
synthesis
and
increase
apoptosis
via
eIF‐2
 phosphorylaFon[5]

Immuno‐suppression


S4
 T.
congolense


T.
evansi
 4
/
23
/
129
31
/
43
/
329
 CalreNculin
precursor
 CalreFculin
binds
to
cell
surface
and
permits
engulfment
of
live
cells[6] Immuno‐suppression,
_Autoimmunity

Y5
 T.
evansi
 7
/
31
/
147
 Neurotransmission
 inhibitor,
 (Hemolysin)
 UNC‐119
is
located
in
neuron
cell
 bodies
and
acts
cell‐autonomously
 to
inhibit
axon
branching[7] Neurological
disorder
 (Anemia)
 References:
 [1]
Holzmuller
P,
Grébaut
P,
PelNer
JB,
Brizard
JP,
Perrone
T,
Gonza?
M,
Bengaly
Z,
Rossignol
M,
Aso
PM,
Vincendeau
P,
Cuny
G,
Boulangé
A,
Frutos
R.
Secretome
of
animal
trypanosomes:
From
a
Standard
Method
toward
New
DiagnosNc
and
TherapeuNc
 Targets.
Ann
N
Y
Acad
Sci.
2008
Dec;1149:337‐42;
[2]
Grébaut
P,
Chuchana
P,
Brizard
JP,
Deme;re
E,
Seveno
M,
Bossard
G,
Jouin
P,
Vincendeau
P,
Bengaly
Z,
Boulangé
A,
Cuny
G,
Holzmuller
P.
IdenNficaNon
of
total
and
differenNally
expressed
excreted‐secreted
 proteins
from
Trypanosoma
congolense
strains
exhibiNng
different
virulence
and
pathogenicity.
Int
J
Parasitol.
2009
Aug;39(10):1137‐50;
[3]
Kaiserman
D,
Bird
PI.
Control
of
granzymes
by
serpins.
Cell
Death
Differ.
2010
Apr;17(4):586‐95;
[4]
Olsson
T,
Bakhiet
 M,
Edlund
C,
Höjeberg
B,
Van
der
Meide
PH,
Kristensson
K.
BidirecNonal
acNvaNng
signals
between
Trypanosoma
brucei
and
CD8+
T
cells:
a
trypanosome‐released
factor
triggers
interferon‐gamma
producNon
that
sNmulates
parasite
growth.
Eur
J
Immunol.
 1991
Oct;21(10):2447‐54;
[5]
Jennings
MD,
Pavi;
GD.
eIF5
has
GDI
acNvity
necessary
for
translaNonal
control
by
eIF2
phosphorylaNon.
Nature.
2010
May
20;465(7296):378‐81;
[6]
Gold
LI,
Eggleton
P,
Sweetwyne
MT,
Van
Duyn
LB,
Greives
MR,
Naylor
SM,
 Michalak
 M,
 Murphy‐Ullrich
 JE.
 CalreNculin:
 non‐endoplasmic
 reNculum
 funcNons
 in
 physiology
 and
 disease.
 FASEB
 J.
 2010
 Mar;24(3):665‐83.
 [7]
 Knobel
 KM,
 Davis
 WS,
 Jorgensen
 EM,
 BasNani
 MJ.
 UNC‐119
 suppresses
 axon
 branching
 in
 C.
 elegans.
 Development.
2001
Oct;128(20):4079‐92.
 Background
and
aims:
 Animal
trypanosomosis
is
a
major
constraint
to
livestock
producNvity
in
the
tropics
and
has
a
significant
impact
on
the
life
of
 millions
of
people.
In
Africa,
South
America
and
south
east
Asia,
the
disease
is
caused
mainly
by
Trypanosoma
congolense
(A),
T.
 evansi
(B),
T.
vivax
and
T.
brucei
brucei.

 Workflow
and
results:
 We
used
2D‐DIGE
and
staNsNcal
differenNal
analysis
(Progenesis
SameSpot
®)
 
coupled
to
Nano
HPLC
ESI‐Q‐ TOF
to
propose
for
the
first
Nme
a
comparaNve
approach
of
the
secretomes
of
T.
congolense
_{and
T.
evansi} clones
exhibiNng
marked
differences
in
their
virulence
and
pathogenicity
profiles.

 A
 

B


The
 extracellular
 posiNon
 of
 trypanosomes
 in
 the
 bloodstream
 of
 their
 host
 (C)
 requires
 consideraNon
 of
 both
 the
 parasite
 and
 its
 naturally
 excreted‐secreted
 factors
 (secretome)
 in
 the
 course
 of
 pathophysiological
 processes
 (anemia,
 cachexia,
 neurological
 disorders).
 We
 therefore
 developed
and
standardised
a
method
to
produce
purified
secretomes
of
African
trypanosomes[1]_{.
 C}

Surprisingly,
 the
 2D‐DIGE‐MS/MS
 analyNcal
 filter
 highlighted
 few
 differenNally
 expressed
 molecules,
 some
 of
 which
 were
 moreover
 idenNfied
 as
 PutaFve


Uncharacterised
 Protein[2]_{.
 Nevertheless
 and
 interesNngly,
 bioinformaNcs
 allowed
 us
 to
 directly
 link
 several
 proteins
 to
 the
 clinical
 disorders
 observed
 in}

trypanosome‐infected
animals
in
the
field.

 Cellular
mechanisms:
 Conclusions
and
perspec8ves:
 This
first
comprehensive
analysis
shows
how
proteomics
is
powerful
in
the
molecular
idenNficaNon
of
differenNally
expressed
trypanosomes
molecules
correlated
 with
either
the
virulence
process
or
exhibiNng
potenNal
properNes
to
induce
pathogenic
dysregulaNon
of
physiological
funcNons.
Moreover,
deciphering
of
the
 molecular
dialogues
and
conflicts
that
govern
host‐parasite
interplay
is
promising
to
define
new
molecular
targets
for
improved
field
diagnosis
and
new
strategies
 of
interference
with
the
infecNous
process
to
fight
against
animal
trypanosomosis.